Posted by
ctrueden on
Oct 08, 2013; 8:36pm
URL: http://imagej.273.s1.nabble.com/Threshold-problems-tp5005085p5005087.html
Hi Leila,
Are you sure ImageJ isn't autoscaling your display differently across your
images?
What does the pixel probe say? Mouse over your data and look at the raw
numbers spit out in the main ImageJ window's status bar.
Also, can you use an auto-threshold algorithm across all your images to
make your analysis reproducible without needing to hardcode a specific
threshold value?
Regards,
Curtis
On Tue, Oct 8, 2013 at 1:56 PM, Leila Alieh <
[hidden email]> wrote:
> Hello!
>
> I developed a macro to count the number of cells positives for a given
> antibody. The cells are selected depending on size, circularity and
> intensity. I found an intensity value good for some images and I run the
> macro. After that I found that the choosen threshold is too high for one
> image, so that to have a reliable valuation for that image I should use
> half of the threshold chosen for the other images, but this latter
> threshold is really too low for the other images and it gives a wrong
> result. I can't understand why this image is so different because the
> intensity of the cells is not so different by eyes (it's a good quality
> image, the ab signal is strong and there are no shadows). I was wondering
> if I did some mistake during the "editing": I had to cut the images (I'm
> interested only in a small area of them) and save them in a tiff format. Is
> it possible that this editing chenged the intensity values?
> Thank you for your help!!!!
>
> Leila
>
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